Method for producing an array for detecting constituents from a biological sample

ABSTRACT

The invention suggests a method of producing an array for the detection of components from a biological sample, wherein the detection molecules are immobilized on one or more supports, said support(s) is/are embedded and subjected to curing, the support is separated into sections, and the sections are applied on another support.

[0001] The invention relates to a method of producing an array for thedetection of components from a biological sample. The method isparticularly suitable in the production of a compact array, because thedetection molecules of the array can be coated on the array with veryhigh density.

[0002] In clinical diagnostics and food analytics, immobilized detectionmolecules, e.g. enzymes and antibodies, have been used as recognitionsubstances for various components in biological samples for severalyears. Where large numbers of such detection molecules are coatedsimultaneously on a single solid support, an array or microarray isconcerned.

[0003] Using arrays, simultaneous investigation of a large number ofsamples is possible with low expenditure of material and work. Forexample, well-known arrays consist of a large number of microscopicspots, each of which may include identical detection molecules, e.g.single-stranded oligonucleotides, cDNA or antibodies, fixed on a solidsupport such as glass or a polymer. When fixing single-strandedoligonucleotides or cDNA, each spot will include a large number ofcopies of a particular sequence which may have 10-2000 bases.

[0004] In detection or analysis, there is an interaction with thesample, forming e.g. a double strand with a complementary sequence—theso-called hybridization. If the spots of the detection molecules includee.g. copies of particular antibodies, analysis or detection proceeds viainteraction with antigens of a biological sample, thereby formingantigen-antibody complexes.

[0005] When using arrays, the density of coated and immobilizeddetection molecules, e.g. DNAs, antibodies and/or receptors, is highlyimportant.

[0006] Medium-density arrays have 100 to 1000 different immobilizeddetection molecules, and the so-called high-density arrays have about10,000 immobilized molecules. Particularly in the production of theso-called high-density chips or arrays, there are some drawbacks withrespect to the capability of mass production. For example, thesedrawbacks involve the complex immobilization procedure and the costlyproduction process. Another drawback in the previous methods ofproducing arrays or microarrays is that the arrays, particularly forclinical uses, cannot be produced with the required reproducibility.That is, arrays for routine investigations coated e.g. with the sameantibodies or receptors vary in their structure and in the density ofcoated immobilized molecules, rendering comparisons between theinvestigations difficult or impossible.

[0007] Various ways of achieving the above objective have been followed.Thus, WO/09607 A1 describes an arrangement of fibrillar supportsembedded in an embedding medium so as to allow production of thinsections perpendicular to the longitudinal axis of the fibrils, whichsections have identical cut surfaces of all fibrils and may serve asarray. However, arranging a large number of fibrils is complex inpractice and in addition, the materials of the fibrils might adverselyaffect the subsequent analysis of biomolecules. The analysis of complexprotein mixtures is difficult to accomplish. To allow imaging of complexdistribution patterns of biomolecules in biological tissues, a tissuearray has been patented (U.S. Pat. No. 4,914,022). Therein, samples ofselected tissues are embedded together in a paraffin block, dissected,and the sections are placed on a support. While the above inventionenables investigations of the distribution patterns in various tissuesections, classification on a molecular level as e.g. in a Western blotfollowing 1D or 2D separation of protein mixtures is not possible.

[0008] The invention therefore is based on the object of providing amethod of producing an array for the detection of components from abiological sample, which method allows for low-cost and simpleproduction of arrays with good reproducibility even with highly complexsamples.

[0009] The present invention solves this technical problem by providinga method of producing an array for the detection of components from abiological sample, wherein the detection molecules are immobilized on afirst support, at least one first support is embedded in a material,said material is subjected to curing, the embedded first support isseparated vertically into sections, at least one section is applied on asecond support such that the detection molecules make contact with thesecond support, and the cured material and/or the first support arecompletely or partially dissolved or removed from the applied sectionusing a solvent, and the array is obtained.

[0010] In the meaning of the invention, arrays can be specific membranecomponents enabling, facilitating or accelerating permeation of materialthrough biomembranes. In the meaning of the invention, however, arraysalso are insoluble organic and inorganic, macromolecular materials suchas porous glass, PVDF membranes and the like, to which solublecomponents such as enzymes and even microorganisms can be bound so as toallow detection of substances via these components.

[0011] In the meaning of the invention, a biological sample is anymaterial obtained from microorganisms, plants, animals, or humans.Biological samples in the meaning of the invention therefore compriseblood, serum, chromosomes, fetuses, tissue samples, gametes, urine,feces, lacrimal fluid, secretions, respiratory air, perspiration, mucoussmears, bone material, cartilage, hair, proteins, nucleotide sequences,carbohydrates, lymph, intestinal contents, degenerate tissue, cells,cell fragments, DNA, RNA, lipids, and the like. Biological samples inthe meaning of the invention can also be subquantities collected fromwaste waters, residues from industrial processes, bogs, or from otherenvironmental fluids. Hence, a sample in the meaning of the invention isany material collected by sampling, or a portion or small amountthereof, the nature of which is to be investigated by physical, chemicaland/or biological means.

[0012] According to the invention, detection molecules are all thosesubstances allowing determination or detection of components from abiological sample. Where antigenic structures are to be investigated ina biological sample, the detection molecules in the meaning of theinvention are antibodies, for example. However, detection molecules canalso be receptors interacting with signal molecules in the biologicalsample. Moreover, detection molecules can also be DNA or RNA sectionshybridizing with DNA and RNA sections in the biological sample. In themeaning of the invention, detection molecules can also bemicroorganisms, cells or cell extracts.

[0013] A support in the meaning of the invention is a material to whichsoluble detection molecules can be bound.

[0014] In the meaning of the invention, immobilization is understood tocomprise all those methods resulting in a restriction of the mobilityand solubility of detection molecules by chemical, biological and/orphysical means. Immobilization can be accomplished using variousmethods, such as binding of detection molecules to each other or to asupport, entrapping in the network of a polymer matrix, or enclosing bymembranes. As a result of immobilization, the detection molecules notonly are made reusable, but also, they can easily be removed subsequentto the process of interaction with the biological sample. They can beused at much higher local concentrations and in continuous flow systems.The detection molecules can be bound to or immobilized on the support bydirect support binding and by crosslinking. According to the invention,support binding or crosslinking particularly proceeds viaions/adsorption or via covalent binding. Crosslinking in the meaning ofthe invention is crosslinking of detection molecules with each other orwith other polymers. Immobilization by inclusion involves inclusion ofthe detection molecules in gel structures or in membranes.

[0015] According to the inventive method of producing an array, thedetection molecules are immobilized on a planar or non-planar firstsupport. The first support can be a membrane, e.g. a nitrocellulosemembrane. Detection molecules can be immobilized on the first supporte.g. by dripping round spots or imprinting lines thereon.

[0016] As first supports, it is possible to use both planar andnon-planar bodies such as spheres, cuboids, or fibrils and bars whichare impregnated with various detection molecules and subsequentlytransferred into an embeddable and dissectable spatial arrangement.

[0017] Numerous ways of immobilizing the detection molecules on thefirst support are known to those skilled in the art. Immobilization isto be effected in such a way that each probe can be related to a definedposition on the support, and that each position on the first support canbe evaluated independently. However, it may also be desired to havepartial or complete overlap of the application points of differentprobes or to apply mixtures of probes. For example, immobilization canbe effected using a method derived from semiconductor technology.Essentially, the detection molecules can be immobilized on the firstsupport in two principally different ways: firstly, an in situ synthesisof the detection molecules at well-defined positions on the firstsupport by successive coupling of monomeric components of synthesis ispossible; secondly, detection molecules previously synthesized orderived from libraries can be placed and immobilized on well-definedpositions of the particularly functionalized support material. To thisend, both spotting and printing procedures can be used. Spotting isunderstood to comprise procedures wherein liquid drops containing thedetection molecules are placed on the support, forming essentially roundspots as a result of surface interactions and drying. However, otherprinting procedures also enable application of detection molecules inwell-defined areas on the surface of the first support, allowing forstable binding of samples to the substrate surface of the detectionmolecules with high coupling efficiency. Similarly, any measure known tothose skilled in the art for immobilizing biomolecules on e.g. columnmaterials can be used to immobilize the detection molecules on the firstsupport.

[0018] For example, selected immobilization procedures are contact tipprinting, ring-and-pin printing, nanoelectric printing andnanopipetting, bubble jet printing, top spot printing, microcontactprinting, micro-fluidic networks methods, photolithographic activationmethod, photoresist lithography, electrochemical focusing and micro-wetprinting. However, comparatively simple sample applicators from thinlayer chromatography or HPLC autosamplers can also be used.

[0019] Using the above procedures, it is possible to obtain a spot sizeranging from less than one micrometer to more than 1000 micrometers.According to the invention, the probe density per cm² may range fromabout 1 to 1 million, for example. The above-mentioned procedures permiteasy handling and high precision of volumes applied, as well as highlyhomogeneous spots which can be applied in a highly paralleled fashion.In particular, the procedures allowing for direct synthesis of detectionmolecules on the first support involve high integration density and asimple combination of immobilization and hybridization. The detectionmolecules, also referred to as probes, can also be prepared by means ofadditional procedures in order to keep the amount of non-specificallysynthesized or bound detection molecules low.

[0020] However, the detection molecules can also be immobilized on thefirst support using a blot procedure. To this end, detection moleculessuch as antibodies, receptors, tissue extracts, microorganisms and thelike are separated by means of electrophoresis. The electrophoreticseparation can be effected using one-dimensional or multi-dimensionalelectrophoresis, particularly 2D electrophoresis.

[0021] The detection molecules separated by electrophoresis aresubsequently transferred either onto the first support or onto amembrane which is placed on the first support in a following step.

[0022] According to the invention, at least one first support having thedetection molecules immobilized thereon is embedded in a material. Avariety of biological and chemical embedding materials are known tothose skilled in the art. The material should be chemically inert to thelargest possible extent, so as to prevent interaction thereof withdetection molecules in a way that would adversely modify the latter.

[0023] However, modification of the detection molecules by interactionwith the material can also be envisaged. Obviously, it is also possibleto do without embedding in those cases where the first support(s) is/areof a dissectable consistency. That is, if the consistency of thesupport(s) allows dissection without any treatment, the support(s)does/do not require embedding.

[0024] For example, the material may consist of saturated hydrocarbonsand can be present in solid state at room temperature. Furthermore, thematerial can be of a nature which allows liquefaction by means ofconventional laboratory methods, e.g. by heating or ultrasonictreatment. Regarding the support to be embedded, with the immobilizeddetection molecules thereon, the material should retain its liquid stateeven at temperatures below 60° C. Above this temperature, some proteinsand peptides or extracts of tissues and/or microorganisms begin toundergo denaturation.

[0025] In another process step of the method according to the invention,the material is made to cure. For example, curing can be effected bysimple cooling of the material or by addition of chemical substanceswhich initiate curing of the material. However, it is also possible tocure the material by physical exposure such as ultrasound or UVradiation or X-rays. Curing of the material should proceed in such a waythat adverse modification of the first support and of the detectionmolecules is avoided. Curing of the material stabilizes the support inits embedded position, thereby allowing e.g. mechanical, chemical orbiological treatment thereof.

[0026] In a further step of the method according to the invention, theembedded first support(s) is/are separated into vertical sections. If,owing to their consistency, the first supports can be separated intosections without any treatment, e.g. by embedding, separation of courseis effected on non-embedded supports. Where the detection molecules areimmobilized on the support in the form of multiple lines, the sectionscan be separated so as to form sections having multiple spots ofimmobilized detection molecules thereon, which represent portions of theseparated lines. Obviously, the individual spots on the section arespaced apart by the same distance as the lines on the first support.

[0027] In a further step of the method according to the invention, atleast one section is placed on a second support, so that the detectionmolecules make contact with the second support. For example, applicationof multiple sections, e.g. side by side, on the second support can beenvisaged. Sections separated from one and the same first support mayhave a virtually identical structure of coated detection molecules. Inparticular, when applying several of these similar sections on one andthe same second support, efficient clinical routine and screeninginvestigations with appropriate options of comparison and control arepossible. One particularly advantageous option of use isdifferential-diagnostic investigations.

[0028] The section or sections are applied on the second support in suchway that the detection molecules are linked with the second support orcontact the second support. Applying the section on the second supportcan be effected by simple lay-up, where adhesion and cohesion forces mayhave a beneficial influence in fixing the section on the second support.For example, this can be achieved by placing the section on a drop ofliquid and heating to stretch the section. However, the use of specialsubstances or devices well-known to those skilled in the art can also beenvisaged in order to apply one or more sections on the second supportand fix them in a desired position. For example, the second support canbe a glass slide held in horizontal position, so that one or moresections can be placed thereon. However, the sections can also be placedon the bottom of a microtest plate or on a membrane, for example.

[0029] According to the method of the invention, the cured materialand/or the first support are completely or partially removed from thesections, thereby obtaining the array. As a result of complete orpartial removal of first support and/or cured material, the detectionmolecules in particular are bound to the second support. For complete orpartial removal of cured material or first support, various solvents areknown to those skilled in the art. The nature of the solvent should besuch that the detection molecules are retained in theirthree-dimensional structure and functionality to the largest possibleextent. More specifically, the solvent can be selected so as to allowstabilization of the detection molecules and, if required, preservationwith highest possible activity under most various conditions of stresseven for months. As a result of dissolving the first support and/or thematerial, the detection molecules are fixed on the second support, sothat repeated use under technical conditions is possible. The detectionmolecules can be bound by the second support via adsorption, ionicbinding or covalent binding. In those cases where the detectionmolecules are enzymes, proteins, nucleic acids, carbohydrates, lipids,etc., this may also take place inside the original microbial, vegetableor animal cell, or in a virus. Binding of the detection molecules to thesecond support allows multiple and continuous use thereof.

[0030] Binding to the support may also proceed via inclusion, as incases where e.g. the second support is a semipermeable membrane in theform of gels, microparticles or fibers. Now, the detection moleculesthus enclosed are separated from the surrounding biological sample by asemipermeable membrane. In particular, spatial fixing by inclusion hasno influence on the activity of the detection molecules. For example,inclusion of detection molecules is also possible in those cases wherethe second supports are sintered glass frits or blotting membranes,thereby allowing concentrating of the detection molecules. For example,such concentrating can be effected by means of capillary blotting usinga liquid flow or by electroblotting. Concentrating or transfer of thedetection molecules can also be effected by dissolving the first supporton the second support. Preferably, such dissolving can be effected insuch a way that the solvent preferably soaks into the second and firstsupports from below in a semi-dry process. However, the solvent can alsobe applied as an aerosol from above, so that first and second supportswill be soaked from above.

[0031] Furthermore, immobilization on the second support is possible bygel inclusion in carragheen, in alginate, or in ENT polymers, byinclusion in polyacrylamide, or on ceramics and on ceramic supports withpolyamine, and/or by means of crosslinking using glutaraldehyde.

[0032] Furthermore, the second support can be designed as anultrafiltration membrane having entrapped detection molecules positioneddownstream or upstream thereof.

[0033] Macroporous supports are preferred in those cases where a largestpossible surface for adsorption or covalent binding is to be achieved.One possible precondition for covalent fixing of detection molecules isthe presence of functional groups on the support. One possibleactivation procedure, e.g. with dextran gels, is reaction withbromocyanogen. According to the chemical nature of the functionalgroups, various types of binding can be formed, e.g. ethers,thio-ethers, esters, etc. Furthermore, coupling procedures for covalentlinkage of detection molecules to agar, agarose and Sephadex supports,and to silanized surfaces of porous glasses are well-known to thoseskilled in the art. Possible changes in the activity of the detectionmolecules can be avoided or reduced by immobilizing the detectionmolecules via spacers to the second support, the option of treatingfirst and second supports identically or differently being known tothose skilled in the art. More specifically, the spacers confer highermobility to the detection molecules, allowing unimpeded contact with thebiological sample. Optional crosslinking of detection molecules by bi-or multifunctional spacer molecules allows immobilization of greateramounts of detection molecules on the second support. For example, thespacers can be added to the solvent used for complete or partial removalof cured material and/or first support. Obviously, spacers can also beused to immobilize the detection molecules on the first support.

[0034] When separating the detection molecules by means of a 2Delectrophoresis, the separated sections may include such spots or not,depending on the position of the individual spots which, in particular,are point-like in shape. For example, if a spot has a diameter of onemillimeter, and sections are separated in a range of one tenth of amillimeter, about 10 sections would include this spot. If each secondsupport is to include all the spots of the first support, thecorresponding sections, after completed dissection of the first spot andbeginning dissection of the next following spot, are immobilized on thesecond support which already has the sections of the preceding spot.Obviously, it might also be envisaged that the second supports do nothave all the spots of the first support, but merely comprise a few spotsof the first support.

[0035] In a preferred embodiment of the invention, at least two firstsupports are embedded in overlap in the material. Such overlapping of atleast two first supports advantageously allows production of highlycompact arrays. For example, two or more first supports includingdetection molecules arranged in line-shape can be positioned one on topof the other. The density of the detection molecules firstly depends onthe spacing of the lines on each first support and secondly on thepacking density of the embedded first supports with respect to eachother. When selecting very thin first supports, e.g. membranes,high-density packing of multiple membranes one on top of the other ispossible. Advantageously, the first supports can be selected in such away that a highest possible number of detection molecules can bearranged on the surface, e.g. by means of a printer. Conveniently,stability requirements to be met by any array are less important becausethe support actually used is the second support. More specifically, thesecond support can be selected according to stability criteria or othercriteria significant e.g. in clinical routine operation.

[0036] For example, 50, 100 or more first supports, e.g. membranes verythin in shape, can be placed one on top of the other and embedded in thematerial. By dense line-shaped or point-like application of a largenumber of detection molecules and by using a large number of firstsupports arranged in overlap one on top of the other, it is possible toachieve a very high density of detection molecules. Thereafter, thefirst supports arranged in overlap are dissected vertically, e.g. in aperpendicular to the line of coated detection molecules. The section nowincludes the detection molecules of the mutually arranged supports, aswell as a few spots of detection molecules arranged on one support at atime, thereby allowing formation of highly compact arrays.

[0037] In another advantageous embodiment of the invention,microorganisms, cells, cell extracts, ligands, antigens, antibodies,receptors, nucleic acids, lectins, proteins, peptides, glycopeptides,carbohydrates, and/or lipids are employed as detection molecules. Themicroorganisms can be living or dead microorganisms, and livingmicroorganisms in the meaning of the invention may concern both growingand resting cells. For example, microorganisms can be immobilized on thefirst support by adhesion and growth. Passive immobilization resultingin microbial growth will form a biofilm on the first support. Apart fromvan-der-Waals forces, other binding forces such as hydrophobicinteractions, hydrogen bridges, and ionic binding contribute to thedevelopment of adhesion. The adhesiveness depends on the chemicalcomposition, overall charge and age of the microorganisms, and on thecharge, composition, and also, on the porosity of the first support.More specifically, glass, titanium dioxide and zirconium dioxide,cellulose, nitrocellulose, nylon, and polyvinyl alcohol membranes can beused as first supports for microorganisms. The microorganisms can alsobe immobilized on the first support by intracellular and intercellularcrosslinking of cells. Conveniently, biopolymers such as polysaccharidesor proteins and also, synthetic polymers, particularly polyacrylamide,are used for inclusion in a polymer matrix to immobilize themicroorganisms, forming e.g. biopolymer beads. These biopolymer beadshaving the cells enclosed therein are fixed either by chelating or bycrosslinking, using e.g. glutardialdehyde. In the inclusion proceduresused to immobilize microorganisms, the pores of the first supportadvantageously are smaller than the microorganisms. Consequently, themicroorganisms will remain in the inclusion, while substrates andproducts from the biological sample can flow in and out. However, themicroorganisms can also be immobilized on the first support byencapsulation in membranes. In the meaning of the invention, it ispossible to distinguish between encapsulation in solid and in liquidmembranes. For example, solid membranes are formed from prefabricatedmembranes, e.g. in membrane reactors such as hollow fiber membranereactors, or the membrane is formed immediately around the cellsuspension, as is the case in microencapsulation, for example. Liquidmembranes are formed by the phase boundary between two non-miscibleliquids, for example. Advantageously, immobilization has only a smalleffect on the functionality or physiological condition of themicroorganisms. For example, immobilized microorganisms offer theadvantage of easier cell removal and cell retention. Another advantageis that enzymes in the microorganisms, being in their naturalmicro-medium, conveniently have higher resistance to pH and temperatureand higher stability in operation. Advantageously, multi-enzymereactions are therefore easier to perform compared to immobilizedenzymes. In particular, this also applies to coenzyme-dependentreactions with in situ regeneration of coenzymes/cofactors.

[0038] The use of immobilized microorganisms is particularlyadvantageous in those cases where (i) the enzymes involved in theconversion of materials are located inside the cell, (ii) the enzymesisolated from the cells are unstable during and subsequent toimmobilization, (iii) the microorganisms do not contain any interferingenzymes, and/or (iv) the enzymes can easily be inactivated or removed,and (v) the substrates and products have low molecular weights.Obviously, the microorganisms can also be immobilized together withenzymes on the first support. Coimmobilization in the meaning of theinvention is immobilization of intact, living or dead cells ofmicroorganisms together with free or immobilized enzymes; for example,immobilized enzymes, together with whole cells, can be entrapped in acommon matrix, or the enzymes can be directly coupled to living cellswhich are subsequently immobilized on the first support. Advantageously,a high percentage of the microorganisms retain their viability. As aresult of coimmobilization of microorganisms and appropriate enzymes, itis possible, among other things, to render substrates fermentable whichotherwise could not be fermented by the organism, and the products thusformed can interact with the detection molecules or with the biologicalsample. In total, coimmobilization can contribute in expanding thespectrum of use of the array. Furthermore, cell extracts and cellcompartments can be immobilized as detection molecules on the firstsupport. Within a cell, numerous types of molecules are confined tospecific areas, i.e., the compartments. In the cell, this is achieved byseparation by membranes impermeable to particular substances. Inaddition, by having compartments in cells or cell extracts, separatereaction volumes are created wherein free diffusion is blocked to alarge extent, because the molecules or enzymes are bound to well-definedstructures. Advantageously, application of cell extracts or cellcompartments will also immobilize the separate reaction volumes on thefirst support.

[0039] Obviously, it is also possible to use ligands as detectionmolecules. For example, ligands in the meaning of the invention aremolecules such as proteins or ions which can be arranged around acentral structure. Ligands can be monodentate and polydentate. However,ligands can also be regarded as molecules that are bound to specificsites of macromolecules, e.g. substrates or coenzymes to a protein.

[0040] Antigens and/or antibodies can also be used as detectionmolecules with advantage. Antigens in the meaning of the invention areall those substances capable of inducing an immune response. These canbe exogenic, natural or synthetic macromolecules, especially proteinsand polysaccharides, with a molecular weight of more than 2 kD, as wellas surface structures of foreign particles. An antigen according to theinvention may comprise a high-molecular weight portion serving assubstrate for—mostly several—low-molecular weight groups, e.g. haptens,which are crucial to the specificity of the immune response and to thereaction of antigens with the corresponding immunoglobulins. Theantigens can be polyvalent and monovalent, thus being capable ofinteracting with one or more types of antibodies.

[0041] However, immobilization of antibodies instead of antigens on thefirst support can also be envisaged. More specifically, antibodies areunderstood to be glycoproteins interacting specifically with an antigen.This interaction results in the formation of antigen-antibody complexes.For example, the antibodies can be various groups of immunoglobulins.Obviously, the antibodies can be immobilized in the form of intactantibodies or in the form of various fragments which can be obtainede.g. by cleavage using various peptidases. The antibodies can bemodified prior to, during or subsequent to immobilization on the firstsupport, e.g. by reduction, oxidation, or by oligomerization.

[0042] Receptors can also be used as detection molecules with advantage.For example, receptors are proteins interacting with extracellularsignal molecules, e.g. with a ligand, activating or initiating specificfunctions via conformational changes, particularly via secondarymessenger substances. However, receptors in the meaning of the inventioncan also be specific cells which receive stimuli and pass on thecorresponding information; examples are photo-, chemo-, thermo- andbaroreceptors.

[0043] Furthermore, lectins can also be immobilized as detectionmolecules on the first support. For example, lectins according to theinvention are sugar-binding proteins or glycoproteins of non-immuneorigin, which agglutinate cells and/or precipitate glycoconjugates.However, proteins may also be concerned, which specifically bind complexsaccharides, but do not agglutinate or precipitate the latter. Forexample, such proteins are monovalent lectins. The lectins can berecovered from plants, invertebrates, vertebrates and/or microorganisms.They can be used as detection molecules to bind e.g. erythrocytes,leukemia cells, yeasts, and some types of bacteria. Advantageously,binding of lectins with substances from a biological sample issaccharide-specific. For this reason, immobilized lectins advantageouslydo not agglutinate those cells lacking the appropriate surfacesaccharides; that is, such cells could be excluded by an infectiousagent or by non-binding. Examples of lectins to be used are concanavalinA from jack beans, agglutinins from wheat germs, lima beans, gardenbeans, soy beans, castor beans, and potatoes. More specifically, it ispossible to detect various types of protein maturing when usingconcanavalin A.

[0044] So-called aptamers can also be used as detection molecules. Inparticular, they are understood to be peptides, DNA or RNA moleculeswhich, owing to their specific molecular shape and charge, are capableof binding other molecules in a specific fashion.

[0045] According to another advantageous embodiment of the invention,the detection molecules are immobilized on the support in the form ofmultiple lines. For example, the first support can be a gel which isused to separate an extract or a mixture of detection molecules.Depending on the type of application on the gel, separate bands orcontinuous lines would form, according to the isoelectric point,molecular weight or the like. However, the detection molecules in thegel can also be blotted on a membrane which then acts as first support.Other methods of immobilizing the detection molecules in multiple lineson a support will be familiar to those skilled in the art. For example,possible procedures are contact tip printing, ring-and-pin printing,photoresist lithography, and micro-wet printing.

[0046] In another advantageous embodiment of the invention, it isenvisaged to transfer the detection molecules from a gel onto the firstsupport by means of Western and/or Southern blotting.

[0047] In another advantageous embodiment of the invention, the use ofsupports comprising nitrocellulose, polyvinylidene difluoride (PVDF),cellulose acetate, cellulose mixed esters, polytetrafluoroethylene(PTFE), polyamide, regenerated cellulose, polycarbonate, polyester,polysulfone, polyacrylamide, agarose, nylon, and/or polyprene as firstsupport is envisaged. In particular, the first support can be selectedsuch that immobilization of a very large number of detection moleculesis possible, because it is the second support that is actually used assupport. Advantageously, the first support especially may comprisenitrocellulose. Nitrocelluloses in the meaning of the invention areinorganic cellulose esters. Furthermore, it is possible to use nylon asfirst support, with nylon in the meaning of the invention comprisinglinear aliphatic polyamides which, in particular, have a high meltingpoint. It is also convenient to use polyvinylidene fluorides as firstsupport. Polyvinylidene fluorides are thermoplastics which are easy toprocess and advantageously, have a high resistance when exposed totemperature and chemicals. Obviously, it is possible to usepolyvinylidene fluorides having varying degrees of crystallinity whichcan be achieved by rapid or slow cooling, particularly duringproduction. Advantageously, polyvinylidene fluorides are remarkable fortheir high mechanical strength, rigidity and tenacity even at lowtemperatures. Conveniently, the first support may also comprisecellulose acetate, with cellulose acetate in the meaning of theinvention being cellulose esters produced by reaction of linters orcellulose with acetic anhydride in acetic acid or methylene chloride assolvent, using strong acids as catalyst in a batch process.Advantageously, cellulose acetates are remarkable for their highstrength, impact strength, scratch resistance, translucency, and surfacegloss, and they can easily be surface-finished by imprinting,varnishing, hot embossing or metallizing. Advantageously, the firstsupport may also comprise cellulose esters or other cellulosederivatives. In the meaning of the invention, cellulose derivatives aresubstances formed by polymer-analogous reactions of chemically modifiedcellulose. Included are products wherein hydroxy hydrogen atoms in theanhydroglucose units of the cellulose are substituted by organic orinorganic groups solely by esterification and/or etherificationreactions, as well as products formed by formal replacement of hydroxygroups in the natural polymer by functional groups not bound via anoxygen atom, or formed via intramolecular elimination of water or byoxidation reactions. In the meaning of the invention, cellulose estersare cellulose derivatives which can be produced by esterification oflinters or cellulose with organic and/or inorganic acids or acidderivatives which also can be used as mixtures. Especially cellulosemixed esters may have good water insolubility and/or thermoplasticproperties. Obviously, the first support may also comprise other plasticmaterials and/or polymers. Plastic materials in the meaning of theinvention include modified natural materials, thermoset materials andthermoplastics, as well as synthetic plastics, e.g. polycondensates,polymers and polyadducts. Examples of such compounds are:acrylonitrile-butadiene-styrene, acrylonitrile-methyl methacrylate,cellulose acetobutyrate, cresol-formaldehyde, carboxymethyl-cellulose,casein, diallyl phthalate, ethylcellulose, epoxide, expandablepolystyrene, ethylene-vinyl acetate, ethylene-vinyl alcohol,tetrafluoroethylene-hexafluoropropylene, high-density polyethylene(rigid PE), low-density polyethylene (plasticized PE), methylmethacrylate-butadiene-styrene, methylcellulose, melamine-formaldehyde,polyamide, polymer of ε-caprolactam, polycondensate ofhexamethyl-enediamine and adipic acid, polyacrylonitrile, polybutene,polybutylene terephthalate, polycarbonate, polychloro-trifluoroethylene,polyethylene, chlorinated polyethylene, ethylene-propylene, polyethyleneterephthalate, phenol-formaldehyde, polyimide, polyisobutylene,polymethyl methacrylate, polyoxymethylene, polyacetal, polypropylene,polyphenylene oxide, polyphenylene sulfide, polystyrene,polytetrafluoroethylene, polyurethane, polyvinyl acetate, polyvinylalcohol, polyvinyl chloride, polyvinylcarbazole, polyvinylpyrrolidone,styrene-acrylonitrile, polystyrene modified with elastomer based onbutadiene, silicone, styrene-α-methylstyrene, urea-formaldehyde, and/orunsaturated polyesters.

[0048] Advantageously, such plastic materials have low specific weight,high resistance to corrosion and high electric insulating capacity, easymolding, and good printability, especially for detection molecules.Polymers in the meaning of the invention are products formed bypolyreactions, e.g. by polymerization, polyaddition and/orpolycondensation, or by polymer-analogous reactions. Monomers resultingin polymer compounds can be e.g. ethylene, styrene, vinyl chloride,vinyl acetate, methyl methacrylate, and/or ethylene oxide.

[0049] In another advantageous embodiment of the invention, the use ofsaturated aliphatic hydrocarbons, especially paraffin, embedding resins,collagen solutions, aqueous solutions, solutions of crosslinkingproteins, carbohydrates, nucleic acids, polymers, and/or agarose ascuring material is envisaged. Hydrocarbons in the meaning of theinvention are organic compounds comprising carbon and hydrogen.Obviously, the hydrocarbons may also comprise further substances orcompounds such as oxygen, nitrogen, sulfur, phosphorus, or specificfunctional groups. Paraffins in the meaning of the invention represent asolid or liquid mixture of especially saturated aliphatic hydrocarbons,which advantageously is colorless, odorless and tasteless, readilysoluble in xylene, ether and chloroform, insoluble in water and 70%alcohol, and does not exhibit fluorescence. Collagen solutions in themeaning of the invention comprise collagen. Collagens are long-fiber,linear-colloidal, high-molecular weight scleroproteins particularlyoccurring in the extracellular matrix, in connective tissue, in theproteinaceous background matrix of bones, and in dentin. In the meaningof the invention, agarose comprises gel-forming polysaccharides fromagar, especially consisting of alternating moieties of β-1,3-linkedD-galactopyranose and α-1,4-linked 3,6-anhydro-L-galactopyranose,wherein the galactose can be methylated in the 6-position. Aqueoussolutions are solutions which comprise water and, in particular, undergocuring at low temperatures.

[0050] In another advantageous embodiment of the invention, the use ofsupports comprising a metal, polypropylene, teflon, polyethylene,polyester, polystyrene, ceramics, and/or glass as second support isenvisaged. Metals in the meaning of the invention are all thosematerials wherein cohesion is furnished by a crystal lattice. Thedividing-line between metals and non-metals is blurred, so that theelements Ce, Sn, As, and Sb are also metals in the meaning of theinvention. Metals according to the invention also include metallicglasses, i.e., materials being in a metastable, largely amorphous state.Obviously, polymers having metallic conductivity are also included inthe meaning of the invention. Advantageously, metals in the meaning ofthe invention have good strength, good hardness and wear resistance,high tenacity, and good electric and thermal conductivity.Polypropylenes in the meaning of the invention are thermoplasticpolymers of propylene. Polypropylenes are remarkable particularly fortheir high hardness, resilience, rigidity, and heat resistance. However,the second support may also comprise teflon. Teflon in the meaning ofthe invention is a polytetrafluoroethylene which advantageously has goodthermoplastic properties. More specifically, polyethylenes are formed bypolymerization of ethylene according to essentially two differentmethods, i.e., the high-pressure and low-pressure processes.Polyethylenes produced in the high-pressure process advantageously havelow density. Essentially, the properties of the second supportscomprising polypropylene are determined by the character of thepolyethylene as a partially crystalline hydrocarbon. Advantageously,polyethylenes are virtually insoluble in all common solvents up to 60°C. Advantageously, polar liquids such as alcohols, esters and ketonesbarely cause swelling of polyethylenes at room temperature.Advantageously, polyethylenes are completely inert when exposed towater, alkaline solutions, salt solutions and inorganic acids. Forexample, supports comprising polyethylenes have a very low water vaporpermeability. Conveniently, the second support may also comprisepolyesters. Polyesters in the meaning of the invention are compoundsproduced by ring-opening polymerization of lactones or bypolycondensation of hydroxycarboxylic acids or of diols and dicarboxylicacids or dicarboxylic acid derivatives. Polyesters in the meaning of theinvention also comprise polyester resins, polyester imides, polyesterrubbers, polyesterpolyols, and polyesterpolyurethanes. Advantageously,polyesters are thermoplastics and have distinct material character. Theyare remarkable for their high thermal stability and can be processedinto alloys with metals such as copper, aluminum and magnesium. However,It can also be envisaged that the second support comprises ceramics.Ceramics in the meaning of the invention is a collective term for anespecially inorganic class of materials predominantly consisting ofnon-metallic compounds and elements and particularly comprising morethan 30% by volume of crystalline materials. Various ceramics or ceramicmaterials which can be used as second support will be familiar to thoseskilled in the art. For example, pottery, earthenware crockery, splitwall tiles, laboratory porcelain, crockery porcelain, bone china,aluminum oxide ceramics, permanent magnet materials, silica bricks, andmagnesia bricks can be concerned. In the meaning of the invention,clay-ceramic materials are classified in coarse and fine materials, withfine clay-ceramic materials comprising earthenware, stoneware andporcelain. Advantageously, specialty ceramics such as glass and oxideceramics, SiC bricks, and melt-cast bricks can also be used as secondsupports. Preferably, the second support may also comprise glass. Glassin the meaning of the invention comprises materials in amorphous,non-crystalline solid state, i.e., the glassy state in the meaning ofthe invention can be regarded as frozen, subcooled liquid or melt. Thus,glass materials are inorganic or organic, mostly oxide melted productsconverted into a solid state by an introduction process withoutcrystallization of the melt phase components. Obviously, crystals,melts, and subcooled melts are also to be regarded as glass materials inthe meaning of the invention. For example, glass materials can be flatglass, container glass, commercial glass, laboratory glass, lead glass,fiber glass, optical fiber glass, and others. obviously, it is alsopossible to use glass materials free of silicate, e.g. phosphate glassmaterials. However, the nature of the second support can be such thatoptical glass, i.e., glass material having a specific optical refractoryindex is used.

[0051] In another advantageous embodiment of the invention, modificationof the support surface is envisaged. More specifically, modification ofthe supports can be effected by biological, physical and/or chemicalexposure. For example, physical exposure would be polishing, etching,pickling, sandblasting, also including physical procedures resulting incuring, coating, finishing, coating with a protective skin, and thelike. For example, surface treatment by biological exposure may includecolonization by microorganisms. Chemical modification of the supportsurface may involve e.g. treatment with acids, bases, metal oxides andothers. The surface of the supports can be modified in such a way thatthe detection molecules have particularly good adherence on the support,or adherence that would not adversely modify the activity thereof.Surface modification also comprises coating with poly-L-lysines,aminosilanes, aldehydesilanes, epoxy groups, gold, streptavidin,branched linkers, reactive groups, polyacrylamide pads, immobilizednitrocellulose, and/or activated aldehydes or agarosealdehyde groups,particularly binding: DNA, COO⁻ groups, NH₂ groups, biotin, thiol groupsand others. Of course, surface modification of the supports alsocomprises a treatment resulting in increased stability and breakingstrength especially of the second support. Obviously, traditionalsurface modification as used in histology, such as coating of first orsecond supports with e.g. proteoglycerol, polylysine, activateddextrans, or bichromated gelatin, can also be performed. Likewise,melt-coating or surface drying of the first support on the secondsupport is possible. Surface drying is a process which preferably canproceed between 20 and 100° C., and between 3720 C. and 80° C. in aparticularly preferred embodiment.

[0052] In another advantageous embodiment of the invention, it isenvisaged to use solvents comprising chloroform, methanol, ethanol, amylacetate, amyl alcohol, cyclohexanone, dimethylsulfoxide,diethylacetamide, dimethylformamide, acetone, acetonitrile, isopropylacetate, methylene chloride, methyl ketone, methyl isobutyl ketone,cellosolve, tetrahydrofuran, methyl acetate, pyridine, butyl acetate,dioxane, ethyl acetate, dimethylacetamide, trifluoroacetic acid,oxidants, acids, bases, and/or enzymes as solvents, particularly inpartial or complete dissolving or removal of the embedding medium or ofthe first support. Solvents in the meaning of the invention aresubstances capable of dissolving others biologically, chemically and/orphysically, particularly inorganic and organic liquids capable ofdissolving other gaseous, liquid or solid substances. For example,inorganic solvents can be classified in solvents containing protons andsolvents free of protons, as well as in aqueous and non-aqueoussolvents. For example, organic solvents are alcohols such as methanol,ethanol, propanols, butanols, octanols, and cyclohexanol, glycols suchas ethylene glycol and diethylglycol, ethers and glycol ethers such asdiethyl ether, dibutyl ether, anisole, dioxane, tetrahydrofuran, mono-,di-, tri-, and/or polyethylene glycol ethers, ketones such as acetone,butanone and cyclohexanone, esters such as acetic acid esters and glycolesters, amides and other nitrogen compounds, such as dimethylformamide,pyridine and acetonitrile, sulfur compounds such as carbon disulfide,dimethylsulfoxide and sulfolane, nitro compounds such as nitrobenzene,halohydrocarbons such as dichloromethane, chloroform,tetrachloromethane, tri- and tetrachloroethene, ethylene chloride, andchlorofluorohydrocarbons, hydrocarbons such as gasolines, petroleumether, cyclohexane, methylcyclohexane, decaline, terpene solvents,benzene, toluene, xylenes, and propylene oxide. However, oxidizingagents can also be used as solvents, e.g. agents which release oxygen totransfer it to other materials, such as potassium permanganate,potassium chlorate and/or lead dioxide, and those having dehydrogenatingeffect, i.e., withdraw hydrogen from other materials or absorb hydrogen,such as iodine and photooxidants; that is, oxidants according to theinvention generally are such elements and compounds having a tendency offorming stable electron shells by electron acceptance to reach a stateof lower energy, e.g. sodium ethanolate, sodium metaperiodate, andhydrogen peroxide. Of course, the solvent may also comprise enzymes.Examples of such enzymes are: oxidoreductases, transferases, hydrolases,lyases, isomerases, and/or ligases, proteases, and collagenase. Enzymesin the meaning of the invention are all those proteins, peptides, lipidsand/or carbohydrates which, as biocatalysts, are capable of modifyingchemical reactions, e.g. cellulase, gelatinase or agarase.

[0053] In another advantageous embodiment of the invention, the possibleuse of a microtome, cryomicrotome, ultramicrotome, ultracryomicrotome,and/or vibratome is envisaged to separate the embedded supports. Toproduce sections, a microtome is particularly suitable in those caseswhere paraffin is used as embedding medium. For example, the outstandingfeature of a cryomicrotome is that, as a result of treatment at lowtemperatures of from −10 to −70° C., the samples are treatedparticularly carefully, and aqueous embedding media can preferably beused with a similar effect of careful treatment. An ultramicrotome isused in those cases where synthetic resins are employed as embeddingmedia and/or particularly thin sections especially with a thickness offrom 20 nm to 1 μm are produced. Such thin sections result in aparticularly economic use of the materials immobilized on the firstsupport. Obviously, the ultramicrotomy can also be performed as acryomicrotomy. In particular, a vibratome is used in those cases wherethe first support to be dissected has a consistency comparable tochemically fixed, biological tissue. In this event, an additionalembedding medium to prepare the sections is not required. A vibratomecan also be used where the embedding medium has appropriate consistency,e.g. serum albumin crosslinked with glutardialdehyde and/or curedcollagen.

[0054] In another advantageous embodiment of the invention, it isenvisaged that detection molecules immobilized on the first support arecrosslinked prior to and/or during and/or subsequent to the removal ofthe first support by incubation with formaldehyde, glutaraldehyde,glacial acetic acid, bivalent coupling reagents, and/or by polyvalentcoupling reagents, particularly oxidized dextran, tosyl-activateddextran, polylysine, activated polylysine, proteins, activated proteins,activated nucleic acids, activated polycarbonates and/or polyethyleneglycols. Incubation with these various substances causes crosslinking ofthe detection molecules in a way to make coating on the second supporteasy.

[0055] One advantage of the method according to the invention is thatseparating the process of immobilization on the first support fromapplying the sections on a second support allows more specific and moreefficient selection of the supports compared to those cases where thesupport having the detection molecules immobilized thereon is the samesupport that is used in detection. In particular, the first support isselected such that a largest possible number of detection molecules canbe immobilized e.g. by existing hydrophobic or hydrophilic interactions,by specific functional groups, by epoxy activation, aldehyde activation,by branched linkers, or by electrically charged macromolecular materialscoated thereon. In contrast, the second support is designed according tospecific aspects of the respective use of the array. For use in clinicaldiagnostics, for example, glass arrays or test strips made of cellulosehave become well-established, but to date, coating and immobilizingdetection molecules thereon has not been possible with the requireddensity, reproducibility and efficiency.

[0056] Another advantage of the method is that the detection moleculesand/or biomolecules coated on the first support can be crosslinked witheach other and, following dissolution of the first support, can form athree-dimensional network on the second support which offers optimumaccess to a component and/or to sample molecules.

[0057] Employing two supports allows the use of e.g. a first supportwith high binding capacity for detection molecules and a second supporthaving good optical properties, i.e., low autofluorescence, for example.By complete or partial dissolution of the first support, it is possibleto reduce or avoid interfering optical properties of the first support.At the same time, however, it is possible to ensure high occupationdensity of detection molecules on the second support, enabling optimumfluorescence detection.

[0058] Another advantage of the method according to the invention isthat the detection molecules can be immobilized with high-densitypacking. In particular, the packing density follows from the spotdensity and/or line density of the detection molecules on a firstsupport and from packing multiple first supports one on top of theother. Using the method of the invention, the arrays can be producedwith high reproducibility and at low cost. By transferring the firstsupport on the bottom of a microtest plate, the detection system isbrought into an automatizable system environment which is standard inmany laboratories and especially in routine laboratories.

[0059] Without intending to be limiting, the invention will be explainedin more detail with reference to the following examples.

EXAMPLE 1 Transfer of a Western Blot from Nitrocellulose on a GlassSlide and Subsequent Determination of Pathogen-Specific Human IgG

[0060] Bacterial crude extracts from Borrelia burgdorferi, Chlamydiatrachomatis, Yersinia enterocolytica, Campylobacter jejuni, Mycoplasmapneumoniae and Salmonella spec. are adjusted to a protein concentrationof 1 mg/ml by diluting with distilled water. Subsequently, 10 μg of eachcrude extract is separated in a polyacrylamide gel disk electrophoresis(Laemmli, 1970, Nature, 227; 680-685) (10% gel, 1 mm gel thickness, comb10, electrophoretic chamber: Mini-Protean II, Biorad). Usingelectroblotting (Towbin et al., 1979, Proc. Natl. Acad. Sci. U.S.A. 76,4350-4354), the separated proteins are transferred from the gel onto anitrocellulose membrane (Life Technologies).

[0061] The dried lanes of each bacterial crude extract are cut out ofthe nitrocellulose film and cut into 20 pieces of 0.5 cm. In awell-defined order and one on top of the other, the pieces are placed ina flexible metal frame which has one interruption and is situated in aglass dish with a planar bottom and a diameter of 5 cm. Between thenitrocellulose pieces having blotted proteins thereon, 3 nitrocellulosepieces of identical size are placed as spacers. The glass dish ispreheated to 60° C. and filled up with liquid paraffin to a height of 1cm without changing the order of the nitrocellulose pieces. Care shouldbe taken to remove air bubbles possibly situated between thenitrocellulose pieces by gentle agitation or by applying a vacuum. Thedish filled with paraffin is incubated at 60° C. overnight. Thereafter,the dish is rapidly cooled, and the metal frame including thenitrocellulose strips is cut out of the whole block. Ultimately, theblock including the nitrocellulose pieces is exposed by bending themetal frame and secured on a microtome in order to prepare the paraffinsections.

[0062] Sections 10 μm in thickness are prepared and placed on glassslides, one at a time, which have been treated with proteoglycerolaccording to standard procedures. Following “stretching” of the paraffinsection on a drop of water at 60° C., the water is sucked off, thesection is positioned and dried at 37° C. for at least about 2 hours.

[0063] Following drying, the sections are made free of paraffin byplacing the slides in xylene. Thereafter, the slides are washed twice in100% ethanol and twice in 100% acetone for 5 minutes each time,subsequently rehydrated in water for 2 minutes each time, andequilibrated in 0.05 M sodium phosphate buffer, pH 7,4, +0.1% Tween, for15 minutes.

[0064] Subsequently, the slides are blocked in 1% human serum albumin inPBS-T for 30 minutes.

[0065] Now, the slides are incubated at room temperature for 1 hour withhuman serums diluted 1:100 in PBS-T. Thereafter, the slides are placedin PBS-T and washed for 30 minutes, changing the washing fluid threetimes. This is subsequently incubated with ScreenBeads-Rhodamin(Chemicell) having anti-human IgG antiserum (IgG fraction) (Rockland)coupled thereto, at a dilution of 1:1000 in PBS-T for 1 hour at roomtemperature. Following washing the slides with PBS-T for 30 minutes,changing the washing fluid three times, and brief immersion in distilledwater, the slides are dried and evaluated using a laser cryptoscope(IOM). The measured rhodamine fluorescence is proportional to the amountof specifically bound human IgG. Alternatively, a peroxidase can becoupled to the anti-human IgG antiserum (IgG fraction). Followingincubation with the antibody-peroxidase conjugate and the subsequentwashing step, the slides are incubated with diaminobenzidine substratesolution according to standard procedures. Evaluation in this case iseffected using a scanner or a CCD camera.

EXAMPLE 2 Transfer of a Western Blot from Nitrocellulose on a PVDFMembrane and Determination of Pathogen-Specific Human IgG

[0066] A bacterial crude extract of Borrelia burgdorferi is separated ina polyacrylamide gel electrophoresis and blotted on a nitrocellulosemembrane as in Example 1. The following specific bands are cut out ofthe nitrocellulose blotting membrane using a scalpel: 83 kD, 41 kD, 39kD, 34 kD, 31 kD (Osp A), 28/29 kD (Osp D), 25 kD (Osp C), 21 kD. Thebands can be composed from different blots to ensure optimumrepresentation of the respective protein. The cut-out bands are thenadhered on a second membrane (nitrocellulose or PVDF) at 2 mm intervalsin a well-defined arrangement. The second membrane now is enclosed in aparaffin block and cut into sections 10 μm in thickness along theperpendicular to the length of the protein bands. The sections arestretched at 56° C. on a water surface, transferred onto a PVDF membraneand dried at 45° C. Subsequently, the paraffin is completely removed byincubation with xylene or Rotihistol (Roth). To this end, the PVDFmembrane is transferred on a filter paper soaked with the solvent andincubated in a sealed chamber for two hours. Thereafter, the PVDFmembrane is incubated twice with 100% methanol in the same way for 5minutes, subsequently in DMSO for 1 hour, and then in water for onehour. Thereafter, the PVDF membrane is equilibrated in 0.05 M Tris-HClbuffer, pH 7.4, and 0.1% Tween 20 (TBS-T) and subsequently blocked forone hour by adding 1% human serum albumin.

[0067] Subsequently, the membranes thus prepared are incubated for onehour with patient serums previously diluted in TBS-T.

[0068] Following several washings in TBS-T, the membranes are incubatedwith anti-human IgG peroxidase (Rockland) at a dilution of 1:2000 inTBS-T for one hour. Following several washings in TBS-T, theperoxidase-substrate reaction is performed for 15 minutes using TMBready-to-use solution (Seramun).

[0069] All incubations in the immune detection are performed at roomtemperature.

1. A method of producing an array for the detection of components from abiological sample, characterized in that the detection molecules areimmobilized on one or more first supports, the first support(s) is/areembedded in a material, said material is caused to cure, the embeddedfirst support(s) is/are separated vertically into sections, at least onesection is applied on a second support, the cured material and/or thefirst support(s) are completely or partially dissolved or removed fromthe applied section using a solvent, and the array is obtained.
 2. Themethod according to claim 1, characterized in that at least two firstsupports are embedded in overlap in the material.
 3. The methodaccording to claim 1 or 2, characterized in that microorganisms, cellextracts, ligands, antigens, antibodies, receptors, nucleic acids,lectins, proteins, peptides, glycopeptides, carbohydrates, and/or lipidsare employed as detection molecules.
 4. The method according to any ofthe preceding claims, characterized in that the detection molecules areimmobilized on the first support in the form of multiple lines.
 5. Themethod according to any of the preceding claims, characterized in thatthe detection molecules are transferred from a gel onto the firstsupport by means of a Western blot and/or Southern blot.
 6. The methodaccording to any of the preceding claims, characterized in that supportscomprising nitrocellulose, polyvinylidene difluoride (PVDF), celluloseacetate, cellulose mixed esters, polytetrafluoroethylene (PTFE),polyamide, regenerated cellulose, polycarbonate, polyester, polysulfone,polyacrylamide, agarose, nylon, and/or polyprene are used as firstsupports.
 7. The method according to any of the preceding claims,characterized in that saturated aliphatic hydrocarbons, especiallyparaffin, embedding resins, collagen solutions, aqueous solutions,solutions of crosslinking proteins, carbohydrates, nucleic acids,polymers, and/or agarose are used as curing material.
 8. The methodaccording to any of the preceding claims, characterized in that supportscomprising a metal, polypropylene, teflon, polyethylene, polystyrene,polyester, ceramics, and/or glass are used as second supports.
 9. Themethod according to any of the preceding claims, characterized in thatthe surfaces of the supports are modified.
 10. The method according toany of the preceding claims, characterized in that solvents comprisingchloroform, methanol, ethanol, amyl acetate, amyl alcohol,cyclohexanone, dimethylsulfoxide, diethylacetamide, dimethylformamide,acetone, acetonitrile, isopropyl acetate, methylene chloride, methylketone, methyl isobutyl ketone, cellosolve, tetrahydrofuran, methylacetate, pyridine, butyl acetate, dioxane, ethyl acetate,dimethylacetamide, trifluoroacetic acid, oxidants, acids, bases, and/orenzymes are used as solvents.
 11. The method according to any of thepreceding claims, characterized in that a microtome, a cryomicrotome, anultramicrotome, an ultracryomicrotome, and/or a vibratome is used toseparate the embedded supports.
 12. The method according to any of thepreceding claims, characterized in that detection molecules immobilizedon the first support are crosslinked with each other prior to and/orduring and/or subsequent to the removal of the first support byincubation with formaldehyde, glutaraldehyde, glacial acetic acid,bivalent coupling reagents, and/or by polyvalent coupling reagents,particularly oxidized dextran, tosyl-activated dextran, polylysine,proteins, activated proteins, activated polylysine, activatedpolycarbonates and/or polyethylene glycols.